保健推拿手法对疲劳型亚健康模型大鼠免疫功能的影响及其NEI机制研究

The health care massage of TCM is often used to relieve the sub-health fatigue, and its disease prevention and health care function may be associated with the immunoregulation. The possible mechanism lies in the following two aspects：(1) The signals produced by the manipulation stimulition pass the peripheral nerve to central nerve,which stimulate central nervous system to produce neuropeptide, neurotransmitter and neuromodulator. All of which can combine regulate cell immunity with the corresponding receptors on immune cells. (2) The functional activities of hypothalamic-pituitary-adrenal (HPA)axis through regulation function of the central nervous system can weaken the immunosuppressive function of the body internal environment , regulating cell immunity. In this study ,60 wister rats are divided into normal feeding group,model group and Chinese massag group. The latter two groups will be treated with partial sleep deprivation,beisdes,the massage group with massage. Before the experiment and during the experiment, We should observe the behavioral changes of the rats on the 7th day and the 14th day,at the same time,experimental specimens will be obtained.Enzyme-linked immunosorbent assay (ELISA) will be used detect the concentration of plasma β-EP、serum corticosterone（CORT）、 adrenocorticotropic hormone(ACTH)、peripheral blood 、 the culture fluid of spleen IL-2、IL-4、IFN-γand so on. After these ,the Flow cytometry is used to detect changes of CD3+、CD4+、CD8+ in the preipheral blood and the apoptosis rate of thymocytes ,and Spleen indexes were determined by gravimetric method.Then light and electron microscopy to observe thymus and spleen morphology,methyl thiazolyl tetrazolium(MTT) colorimetry was used to detect the abilities of spleen lymphocyte proliferation,RT-PCR technology to detect pituitary β-EPmRNA,immunohistochemical Strept Avidin-Biotin Complex（SABC）method to detect hypothalamic corticotropin-releasing hormone and the expression of Spleen lymphocyte Surface.

用于缓解亚健康疲劳的中医保健推拿，其防病保健功能可能与免疫调节相关，其调节机制可能在于：（1）手法经外周神经传入途径将信号传入中枢，刺激中枢神经系统产生神经肽、神经递质与调质，与免疫细胞上相应受体结合从而调节免疫。（2）经中枢调节下调HPA轴功能活动，减弱机体内环境免疫张力性抑制作用，从而调节免疫。 拟将60只大鼠随机分为正常组、模型组和推拿组，后两组给予部分睡眠剥夺，推拿组给予推拿手法。实验前及第7、14d观测一般情况及行为学，第14d取材，ELISA法测垂体及血浆β-EP、血清CORT与ACTH、外周血与脾培养液IL-2与IL-4及IFN-γ；流式细胞术测外周血CD3+、CD4+、CD8+；称重法测脾指数；流式细胞术测胸腺细胞凋亡率；镜下测胸腺及脾脏组织形态学；MTT比色法测脾淋巴细胞增殖；RT-PCR法测垂体β-EPmRNA；SABC法测下丘脑CRH及脾淋巴细胞表面β-EPR表达。

背景：临床观察表明，中医保健推拿对亚健康疲劳具有保健效果，但其机制尚未明确。 目的：观察保健推拿手法对疲劳型亚健康模型大鼠免疫功能的影响及其神经-内分泌-免疫机制。方法：60只大鼠随机分为正常组、模型组和推拿组，后两组给予部分睡眠剥夺，推拿组给予推拿手法。实验前及第7、14d观测一般情况及行为学，第14d取材，ELISA法测垂体及血浆β-EP、血清CORT与ACTH、外周血与脾培养液IL-2与IL-4及IFN-γ；流式细胞术测外周血CD3+、CD4+、CD8+；称重法测脾指数；流式细胞术测胸腺细胞凋亡率；镜下测胸腺及脾脏组织形态学；MTT比色法测脾淋巴细胞增殖；RT-PCR法测垂体β-EPmRNA；SABC法测下丘脑CRH及脾淋巴细胞表面β-EPR表达。结果：（1）模型组出现疲劳型亚健康症状。与正常组比较，模型组血清ACTH、CORT浓度及胸腺细胞早期凋亡率增高，胸腺和脾脏器官质量指数下降，脑垂体β-EP mRNA及下丘脑CRH表达下调，胸腺和脾脏的结构与功能明显损伤。（2）推拿组疲劳型亚健康症状介于正常组与模型组之间。到实验结束，与模型组比较，推拿组血清ACTH、CORT浓度及胸腺细胞早期凋亡率显著下降，丝裂原ConA增殖反应能力增高，外周血T细胞亚群数量增加，胸腺和脾脏器官质量指数提高。（3）与模型组比较，推拿组脑垂体β-EP mRNA及下丘脑CRH表达增加。（4）与模型组比较，推拿组对胸腺与脾脏损伤呈现明确的拮抗作用。结论：用于缓解亚健康疲劳的中医保健推拿，其防病保健功能可能与免疫调节相关，其调节机制可能在于：（1）手法经外周神经传入途径将信号传入中枢，刺激中枢神经系统产生神经肽、神经递质与调质，与免疫细胞上相应受体结合从而调节免疫。（2）经中枢调节下调HPA轴功能活动，减弱机体内环境免疫张力性抑制作用，从而调节免疫。